Quinuclidine carbonate salts and medicinal composition thereof

ABSTRACT

Quinuclidine carbonate derivatives act as muscarinic receptor antagonists and are effective for the prevention and/or treatment of a broncho-obstructive or inflammatory diseases.

CROSS REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.12/512,262, filed on Jul. 30, 2009, now U.S. Pat. No. 8,039,483 andclaims priority to European Patent Application No. 08162066.8 filed onAug. 8, 2008.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to quinuclidine carbonate salts which actas muscarinic receptor antagonists. The present invention also relatesto methods of preparing such a salt, compositions which contain such asalt, and methods of treating certain conditions by administering such asalt.

2. Discussion of the Background

Quaternary ammonium salts which act as muscarinic (M) receptorsantagonists drugs are currently used in therapy to inducebronchodilation for the treatment of respiratory diseases. Examples ofwell known M receptor antagonists drugs are represented by ipratropiumbromide and tiotropium bromide.

Several chemical classes acting as selective M3 receptors antagonistsdrugs have been developed for the treatment of inflammatory orobstructive airway diseases such as asthma and chronic obstructivepulmonary disease (COPD). Quinuclidine carbamate derivatives and theiruse as M3 antagonists are described in WO 02051841, WO 03053966, and WO2008012290.

Said M and M3 receptors antagonists drugs are currently administeredthrough inhalation in order to deliver the drug directly at the site ofaction and hence limiting the systemic exposure. However, even thoughthe inhalatory route limits the systemic exposure, the known compoundsmay still exhibit undesired side effects due to systemic absorption.

It is hence highly desirable to provide M3 receptor antagonists drugsable to act locally, while having high potency and long duration ofaction, and said drugs, once adsorbed, are degraded to inactivecompounds which are deprived of any systemic side effects typical ofmuscarinic antagonists.

SUMMARY OF THE INVENTION

Accordingly, it is one object of the present invention to provide novelM3 receptor antagonists drugs.

It is another object of the present invention to provide novel M3receptor antagonists drugs which act locally.

It is another object of the present invention to provide novel M3receptor antagonists drugs which exhibit a high potency and longduration of action.

It is another object of the present invention to provide novel M3receptor antagonists drugs which, once adsorbed, are degraded toinactive compounds which are deprived of any systemic side effectstypical of muscarinic antagonists.

It is another object of the present invention to provide novel methodsof making such a M3 receptor antagonist.

It is another object of the present invention to provide novelcompositions which contain such a M3 receptor antagonist.

It is another object of the present invention to provide novel methodsof treating certain conditions or diseases by administering such a M3receptor antagonist.

These and other objects, which will become apparent during the followingdetailed description, have been achieved by the inventors' discovery ofquinuclidine carbonate salts represented by formula (I):

wherein:

A is optionally substituted aryl or heteroaryl or arylalkyl orheteroarylalkyl or a group represented by formula (a):

wherein

R₃ and R₄ are the same or different and may be independently selectedfrom the group consisting of H, (C₃-C₈)-cycloalkyl, aryl or heteroaryl,wherein said aryl or heteroaryl may be optionally substituted with ahalogen atom or with one or several substituents independently selectedfrom the group consisting of OH, O—(C₁-C₁₀)-alkyl, oxo (═O), SH,S—(C₁-C₁₀)-alkyl, NO₂, CN, CONH₂, COOH, (C₁-C₁₀)-alkoxycarbonyl,(C₁-C₁₀)-alkylsulfanyl, (C₁-C₁₀)-alkylsulfinyl, (C₁-C₁₀)-alkylsulfonyl,(C₁-C₁₀)-alkyl and (C₁-C₁₀)-alkoxyl or when R₃ and R₄ are bothindependently aryl or heteroaryl they may be linked through a Y groupwhich may be a (CH₂)_(n) with n=0-2, wherein when n=0 Y is a singlebond, forming a tricyclic ring system wherein the carbon atom of(CH₂)_(n) may be substituted by a heteroatom selected from O, S, N andwith the proviso that R₃ and R₄ are never both H;

R is a residue selected from (C₁-C₁₀)-alkyl, (C₂-C₁₀)-alkenyl, and(C₂-C₁₀)-alkynyl optionally substituted with a group selected from:

-   -   NH₂, NR₁R₂, CONR₁R₂, NR₂COR₁, OH, SOR₁, SO₂R₁, SH, CN, NO₂, an        alicyclic compound, Z—R₁ (wherein Z is selected from CO, O, COO,        OCO, SO₂, S, SO, COS and SCO or it is a bond), and        (C₃-C₈)-cycloalkyl, wherein        -   R₁ is a residue selected from:            -   an alicyclic compound optionally substituted with one or                several substituents independently selected from OH, oxo                (═O), SH, NO₂, CN, CONH₂, NR₂CO—(C₁-C_(x))-alkyl, COOH,                (C₁-C₁₀)-alkoxycarbonyl, (C₁-C₁₀)-alkylsulfanyl,                (C₁-C₁₀)-alkylsulfinyl, (C₁-C₁₀)-alkylsulfonyl,                (C₁-C₁₀)-alkyl, and (C₁-C₁₀)-alkoxyl                NR₂CO—(C₁-C₁₀)-alkyl;            -   aryl optionally substituted with NR₂CO—(C₁-C₁₀)-alkyl,                (C₁-C₁₀)-alkyl, O—(C₁-C₁₀)-alkyl or halogen, and            -   heteroaryl optionally substituted with                NR₂CO—(C₁-C₁₀)-alkyl or halogen, wherein        -   R₂ is a group selected from H, phenoxycarbonyl,            benzyloxycarbonyl, (C₁-C₁₀)-alkoxycarbonyl,            (C₁-C₁₀)-alkylcarbonyl, (C₁-C₁₀)-alkylsulfonyl, and            (C₁-C₁₀)-alkyl; and            X⁻ is a physiologically acceptable anion.

The present invention also provides pharmaceutical compositions whichcontain a salt represented by formula (I) alone or in combination withor in mixture with one or more pharmaceutically acceptable carriersand/or excipients.

The present invention also provides the use of a salt represented byformula (I) for preparing a medicament.

In a further aspect, the invention provides the use of a saltrepresented by formula (I) for the prevention and/or treatment of anybroncho-obstructive or inflammatory disease, preferably asthma orchronic bronchitis or chronic obstructive pulmonary disease (COPD).

In a further aspect, the present invention provides the use of a saltrepresented by formula (I) for the manufacture of a medicament for theprevention and/or treatment of any broncho-obstructive or inflammatorydisease, preferably asthma or chronic bronchitis or chronic obstructivepulmonary disease (COPD).

The present invention further provides a method for prevention and/ortreatment of any broncho-obstructive or inflammatory disease, preferablyasthma or chronic bronchitis or chronic obstructive pulmonary disease(COPD), which comprises administering to a subject in need thereof atherapeutically effective amount of a salt represented by formula (I).

The present invention also provides pharmaceutical compositions suitablefor administration by inhalation.

Inhalable preparations include inhalable powders, propellant-containingmetering aerosols or propellant-free inhalable formulations.

The present invention is also directed to devices which may be a single-or multi-dose dry powder inhaler, a metered dose inhaler and a soft mistnebulizer and which contain a salt represented by formula (I).

The present invention is also directed to a kit comprising apharmaceutical composition which contains a salt represented by formula(I) alone or in combination with or in admixture with one or morepharmaceutically acceptable carriers and/or excipients and a devicewhich may be a single- or multi-dose dry powder inhaler, a metered doseinhaler and a soft mist nebulizer and which contains a salt representedby general formula (I).

Thus, the present invention provides quinuclidine carbonate salts withtherapeutically desirable characteristics. The salts represented bygeneral formula (I) behave as soft-drugs, since they are able to producea persistent bronchodilating effect in the lung but are consistently andrapidly transformed into inactive metabolites after passing into humanplasma. This behaviour gives great advantages in terms of safety.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The term “halogen atoms” as used herein includes fluorine, chlorine,bromine, and iodine.

The expression “(C₁-C₁₀)alkyl”, refers to straight-chained and branchedalkyl groups wherein the number of carbon atoms is from 1 to 10.Examples of groups are methyl, ethyl, n-propyl, isopropyl, t-butyl,pentyl, hexyl, heptyl, octanyl, nonenyl, and decenyl.

Optionally, one or more hydrogen atoms in said groups can be replaced byhalogen atoms. The derived expressions “(C₁-C₁₀)-alkoxycarbonyl”,“(C₁-C₁₀)-alkylsulfanyl”, “(C₁-C₁₀)-alkylsulfinyl”,“(C₁-C₁₀)-alkylsulfonyl” and “(C₁-C₁₀)-alkoxyl” are to be construed inan analogous manner.

The derived expressions “(C₂-C₁₀) alkenyl” and “(C₂-C₁₀) alkynyl”, areto be construed in an analogous manner.

The expression “an alicyclic compound” includes:

-   -   “(C₃-C₈)-cycloalkyl”, which refers to cyclic non-aromatic        isolated hydrocarbon saturated groups. Examples include        cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,        and cyclooctenyl;    -   “bicycloalkyl” and “tricycloalkyl” groups which are non-aromatic        saturated cyclic alkyl moieties consisting of two or three        rings, respectively, wherein said rings share at least one        carbon atom. For purposes of the present invention, and unless        otherwise indicated, bicycloalkyl groups include spiro rings and        fused rings. Examples of bicycloalkyl groups include, but are        not limited to, bicyclo-[3.1.0]-hexyl,        bicyclo-[2.2.1]-hept-1-yl, norbornyl, spiro[4.5]decyl,        spiro[4.4]nonyl, Spiro[4.3]octyl, and spiro[4.2]heptyl. An        example of a tricycloalkyl group is adamantanyl; and    -   cyclic non-aromatic hydrocarbon unsaturated groups. Examples        include cyclohexenyl, norbornenyl, bicyclo[2.2.1.]heptanyl.

Optionally, one or more hydrogen atoms in said groups can be replaced byone or more halogen atoms.

The expression “aryl” refers to mono, bi- or tricyclic ring systemshaving 5 to 20, preferably from 5 to 15, ring atoms, and wherein atleast one ring is aromatic. Optionally, one or more hydrogen atoms insaid rings can be replaced by one or more halogen atoms or phenyl.

The expression “heteroaryl” refers to mono, bi- or tricyclic ringsystems having 5 to 20, preferably from 5 to 15, ring atoms, in which atleast one ring is aromatic and in which at least one ring atom is aheteroatom (e.g. N, S, or O). Optionally, one or more hydrogen atoms insaid rings can be replaced by one or more halogen atoms.

The expression “arylalkyl” refers to a “(C₁-C₄) alkyl” optionallysubstituted by a mono, bi- or tricyclic ring systems which have 5 to 20,preferably from 5 to 15, ring atoms. Optionally, one or more hydrogenatoms in said rings can be replaced by one or more halogen atoms.

Examples of suitable arylalkyl groups include benzyl, biphenylmethyl andthiophenylmethyl.

The expression “heteroarylalkyl” refers to a “(C₁-C₄) alkyl” optionallysubstituted by a mono, bi- or tricyclic ring systems which have 5 to 20ring atoms, preferably from 5 to 15, in which at least one ring isaromatic and in which at least one ring atom is a heteroatom (e.g. N, Sor O). Optionally, one or more hydrogen atoms in said rings can bereplaced by one or more halogen atoms.

Examples of suitable monocyclic systems include thiophene,cyclopentadiene, benzene, pyrrole, pyrazole, imidazole, isoxazole,oxazole, isothiazole, thiazole, pyridine, imidazolidine, piperidine, andfuran radicals.

Examples of suitable bicyclic systems include naphthalene, biphenyl,purine, pteridine, benzotriazole, quinoline, isoquinoline, indole,isoindole, and benzothiophene radicals.

Examples of suitable tricyclic systems include fluorene radicals.

The invention is directed to quinuclidine carbonate salts which act asmuscarinic receptors antagonists, said salts preferably acting on the M3receptors.

Advantageously, physiologically acceptable anions X include thoseselected from the group consisting of chloride, bromide, iodide,sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate,citrate, fumarate, tartrate, oxalate, succinate, benzoate, andp-toluenesulfonate, preferably chloride, bromide and iodide, morepreferably chloride and bromide.

A preferred group of salts of formula (I) is that wherein R is(C₁-C₆)-alkyl substituted by —Z—R₁, wherein Z and R₁ are as describedabove.

Another preferred group of salts of formula (I) is that wherein R is(C₁-C₆)-alkyl substituted by —Z—R₁, wherein Z is O, CO or a bond and R₁is aryl or heteroaryl as described above, preferably substituted withone or more halogen atoms.

A more preferred group of salts of formula (I) is that wherein R is CH₂,Z is CO and R₁ is thienyl, according to formula (II):

wherein A is as defined above.

A first group of salts of formula (II) is represented by the saltswherein A is optionally substituted aryl or heteroaryl or arylalkyl orheteroarylalkyl or a group of formula (a)

wherein R₃ and R₄ are both independently aryl or heteroaryl, preferablysubstituted with one or more halogen atoms.

A second group of salts of formula (II) is represented by the saltswherein A is a group of formula (a), wherein R₃ and R₄ are both phenyl,preferably substituted with one or more halogen atoms.

A third group of salts of formula (II) is represented by the saltswherein A is a compound of formula (a), wherein R₃ and R₄ are bothindependently phenyl and they are linked through a Y group which may bea (CH₂)_(n) with n=0-2, wherein when n=0, Y is a single bond, forming atricyclic ring system of formula (b), wherein the carbon atom of(CH₂)_(n) may be substituted by a heteroatom selected from O, S, N.

A fourth group of salts of formula (II) is represented by the saltswherein A is a 9H-fluoren-9-yl, preferably substituted with one or morehalogen atoms.

A fifth group of salts of formula (II) is represented by the saltswherein A is an arylalkyl, preferably a phenyl-(C₁-C₄)-alkyl, morepreferably substituted with one or more halogen atoms and even morepreferably A is a benzyl, optionally substituted with one or morehalogen atoms.

A sixth group of salts of formula (II) is represented by the saltswherein A is a biphenylmethyl, preferably substituted with one or morehalogen atoms.

A seventh group of salts of formula (II) is represented by the saltswherein A is a thiophenylmethyl, preferably substituted with one or morehalogen atoms.

Another preferred group of salts of formula (I) is that wherein R is apropyl substituted by —Z—R₁ group wherein Z is O and R₁ is phenyl,according to formula (III):

wherein A is as defined above.

A first group of salts of formula (III) is represented by the saltswherein A is optionally substituted aryl or heteroaryl or arylalkyl orheteroarylalkyl or a group of formula (a)

wherein R₃ and R₄ are both independently aryl or heteroaryl, preferablysubstituted with one or more halogen atoms.

A second group of salts of formula (III) is represented by the saltswherein A is a group of formula (a), wherein R₃ and R₄ are both phenyl,preferably substituted with one or more halogen atoms.

A third group of salts of formula (III) is represented by the saltswherein A is a group of formula (a), wherein R₃ and R₄ are both phenyland they are linked through a Y group which may be a (CH₂)_(n) withn=0-2, wherein when n=0, Y is a single bond, forming a tricyclic ringsystem of formula (b), wherein the carbon atom of (CH₂)_(n) may besubstituted by a heteroatom selected from O, S, N.

A fourth group of salts of formula (III) is represented by the saltswherein A is a 9H-fluoren-9-yl, preferably substituted with one or morehalogen atoms.

A fifth group of salts of formula (III) is represented by the saltswherein A is arylalkyl, preferably a phenyl-(C₁-C₄)-alkyl, morepreferably substituted with one or more halogen atoms and even morepreferably A is a benzyl, optionally substituted with one or morehalogen atoms.

A sixth group of salts of formula (III) is represented by the saltswherein A is a biphenylmethyl, preferably substituted with one or morehalogen atoms.

A seventh group of salts of formula (III) is represented by the saltswherein A is a thiophenylmethyl, preferably substituted with one or morehalogen atoms.

Another preferred group of salts of formula (I) is that wherein R is amethyl substituted by —Z—R₁ group wherein Z is CO and R₁ is phenyl,according to formula (IV):

Another preferred group of salts of formula (I) is that wherein R is amethyl substituted by —Z—R₁ group wherein Z is CO and R₁ is thienyl,according to formula (V):

It will be apparent that the salts of formula (I) may contain asymmetriccenters. Therefore the invention also includes the optical stereoisomersand mixtures thereof.

Where the compounds according to the invention have at least oneasymmetric center, they may exist as enantiomers. Where the compoundsaccording to the invention possess two or more asymmetric centers, theymay additionally exist as diastereoisomers. It is to be understood thatall such isomers and mixtures thereof in any proportion are encompassedwithin the scope of the present invention.

The active salts (I) shows at least one chiral center, which isrepresented by the quinuclidine carbon atom bearing the carbonate group.

In the preferred embodiments, the active compound (I) is in the form ofthe substantially pure (R)-enantiomer, wherein the enantiomeric purityis higher than 85%, more preferably than 90%, more preferably than 95%and even more preferably than 99%.

According to specific embodiments, the present invention provides thecompounds reported below:

Compound Chemical name 1(R)-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 2(R)-3-Benzhydryloxycarbonyloxy-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 3(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 4(R)-3-(3-Fluoro-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 5(R)-3-(4-Fluoro-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 6(R)-3-(4-Bromo-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride; chloride 7(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 8(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-phenethyl-1-azonia-bicyclo[2.2.2]octane; bromide 9(R)-1-Benzyl-3-(9H-fluoren-9-yloxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane; chloride 10(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-methyl-1-azonia-bicyclo[2.2.2]octane; iodide 11(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-(3-phenoxy-propyl)-1-azonia-bicyclo[2.2.2]octane; bromide 12(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-(3-phenoxy-propyl)-1-azonia-bicyclo[2.2.2]octane; bromide 13(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-methyl-1-azonia-bicyclo[2.2.2]octane; iodide 14(R)-1-(2-Oxo-2-thiophen-2-yl-ethyl)-3-(thiophen-2-ylmethoxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane; chloride 15(R)-3-[Bis-(4-chloro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 16(R)-3-[Bis-(4-bromo-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 17(R)-3-[Bis-(4-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 18(R)-3-(3-Bromo-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 19(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-phenyl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 20(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-carbamoylmethyl-1-azonia-bicyclo[2.2.2]octane; bromide 21(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-p-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 22(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(4-fluoro-phenyl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane; bromide 23(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(4-methoxy-phenyl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane; bromide 24(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-3-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 25(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-phenylcarbamoylmethyl-1-azonia-bicyclo[2.2.2]octane; bromide 26(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(5-chloro-thiophen-2-yl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane; bromide 27(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(3,5-dibromo-thiophen-2-yl)-2-oxo-ethyl]-1-azonia- bicyclo[2.2.2]octane;chloride 28 (R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiazol-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 29(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-o-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 30(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-m-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 31(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(4-methyl-pent-3-enyl)-1-azonia-bicyclo[2.2.2]octane; bromide 32(R)-1-(2-Benzo[b]thiophen-2-yl-2-oxo-ethyl)-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-azonia-bicyclo[2.2.2]octane; bromide 33(R)-1-Benzyl-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-azonia-bicyclo[2.2.2]octane; bromide 34(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-phenoxy-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 35(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-methyl-1-azonia-bicyclo[2.2.2]octane; iodide 36(R)-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-pyridin-4-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; bromide 37(R)-3-[(2-Fluoro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride38 (R)-3-[Bis-(2-chloro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 39(R)-1-(2-Oxo-2-thiophen-2-yl-ethyl)-3-(phenyl-o-tolyl-methoxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane; chloride 40(R)-3-[(3-Fluoro-phenyl)-(3-methoxy-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 41(R)-3-[Cyclohexyl-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 42(R)-3-[(3-Chloro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride43 (R)-3-[(3,5-Difluoro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 44(R)-3-[(3-Fluoro-phenyl)-m-tolyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 45(R)-3-[(3-Fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 46(R)-3-[(3-Fluoro-phenyl)-(4-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride47 (R)-3-[(3,4-Difluoro-phenyl)-phenyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride 48(R)-3-[(3-Fluoro-phenyl)-phenyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane; chloride

The salts of formula (I) may be prepared according to known methods.Some of the processes which can be used are described below and reportedin Scheme 1.

General Procedure for the Preparation of Salts of Formula (I).

The salts of general formula (I) may be prepared according to a generalprocess which includes the following steps:

1^(st) step—An amino-alcohol of formula (2) is reacted with1,1′-carbonyldiimidazole (1) in an aprotic solvent. Advantageously, thereaction is carried out in a solvent selected from CH₃CN, CH₂Cl₂, CHCl₃,tetrahydrofuran (THF), and dioxane at a temperature ranging from 0° C.to the boiling point of the solvent. Water is added, and the imidazolederivative (3) is extracted with a solvent. Examples of solvents whichcan be suitably used are ethyl acetate, diethyl ether, methylenechloride;

2^(nd) step—the solvent is evaporated to dryness, and the residue isreacted with an alcohol of formula (4) in an aprotic solvent.Advantageously, the solvent is selected from THF, dimethylformamide(DMF), and dimethylacetal (DMA). Preferably, the alcohol is activatedwith a base advantageously selected from NaH, BuLi (butyl lithium), andlithium diisopropylamide (LDA), to give a compound of formula (5).

3^(rd) step—Compounds of formula (5) may be alkylated at the nitrogenatom of the tertiary amine by means of an alkylating agent RX wherein Ris as previously defined and X is, according to known conditions,chlorine, bromine, or iodine, giving a quaternary ammonium salt offormula (I).

The invention also provides pharmaceutical compositions of salts offormula (I) in admixture with one or more pharmaceutically acceptablecarriers, for example those described in Remington's PharmaceuticalSciences Handbook, XVII Ed., Mack Pub., N.Y., U.S.A.

Administration of the compounds of the present invention may beaccomplished according to patient needs, for example, orally, nasally,parenterally (subcutaneously, intravenously, intramuscularly,intrasternally and by infusion), by inhalation, rectally, vaginally,topically, locally, transdermally, and by ocular administration.

Various solid oral dosage forms can be used for administering compoundsof the invention including such solid forms as tablets, gelcaps,capsules, caplets, granules, lozenges and bulk powders. The compounds ofthe present invention can be administered alone or combined with variouspharmaceutically acceptable carriers, diluents (such as sucrose,mannitol, lactose, starches) and known excipients, including suspendingagents, solubilizers, buffering agents, binders, disintegrants,preservatives, colorants, flavorants, lubricants and the like. Timerelease capsules, tablets and gels are also advantageous inadministering the compounds of the present invention.

Various liquid oral dosage forms can also be used for administeringcompounds of the invention, including aqueous and non-aqueous solutions,emulsions, suspensions, syrups, and elixirs. Such dosage forms can alsocontain suitable known inert diluents such as water and suitable knownexcipients such as preservatives, wetting agents, sweeteners,flavorants, as well as agents for emulsifying and/or suspending thecompounds of the invention. The salts of the present invention may beinjected, for example, intravenously, in the form of an isotonic sterilesolution. Other preparations are also possible.

Suppositories for rectal administration of the compounds of theinvention can be prepared by mixing the compound with a suitableexcipient such as cocoa butter, salicylates and polyethylene glycols.

Formulations for vaginal administration can be in the form of cream,gel, paste, foam, or spray formula containing, in addition to the activeingredient, such as suitable carriers, are also known.

For topical administration, the pharmaceutical composition can be in theform of creams, ointments, liniments, lotions, emulsions, suspensions,gels, solutions, pastes, powders, sprays, and drops suitable foradministration to the skin, eye, ear or nose. Topical administration mayalso involve transdermal administration via means such as transdermalpatches.

For the treatment of the diseases of the respiratory tract, the saltsaccording to the invention are preferably administered by inhalation.

Inhalable preparations include inhalable powders, propellant-containingmetering aerosols or propellant-free inhalable formulations.

For administration as a dry powder, single- or multi-dose inhalers knownfrom the prior art may be utilized. In that case, the powder may befilled in gelatine, plastic or other capsules, cartridges or blisterpacks or in a reservoir.

A diluent or carrier, generally non-toxic and chemically inert to thesalts of the present invention, e.g. lactose or any other additivesuitable for improving the respirable fraction may be added to thepowdered compounds of the invention.

Inhalation aerosols containing propellant gas such as hydrofluoroalkanesmay contain the salts of the invention either in solution or indispersed form. The propellant-driven formulations may also containother ingredients such as co-solvents, stabilizers and optionally otherexcipients.

The propellant-free inhalable formulations comprising the salts of theinvention may be in form of solutions or suspensions in an aqueous,alcoholic or hydroalcoholic medium and they may be delivered by jet orultrasonic nebulizers known from the prior art or by soft-mistnebulizers such as Respimat®.

The salts of the invention may be administered as the sole active agentor in combination with other pharmaceutical active ingredients includingthose currently used in the treatment of respiratory disorders, e.g.beta₂-agonists, corticosteroids, and PDE4 inhibitors.

The dosages of the salts of the invention depend upon a variety offactors including the particular disease to be treated, the severity ofthe symptoms, the route of administration, the frequency of the dosageinterval, the particular compound utilized, the efficacy, toxicologyprofile, and pharmacokinetic profile of the compound.

Advantageously, the salts of formula (I) can be administered forexample, at a dosage comprised between 0.001 and 1000 mg/day, preferablybetween 0.1 and 500 mg/day.

When the salts of formula (I) are administered by inhalation route, theyare preferably given at a dosage comprised between 0.001 and 500 mg/day,preferably between 0.1 and 200 mg/day.

The salts of formula (I) may be administered for the prevention and/ortreatment of broncho-obstructive or inflammatory diseases, such asasthma, chronic bronchitis, chronic obstructive pulmonary disease(COPD), bronchial hyperreactivity, cough, emphysema or rhinitis;urological disorders such as urinary incontinence, pollakiuria,cystospasm, chronic cystitis and overactive bladder (OAB);gastrointestinal disorders such as bowel syndrome, spastic colitis,diverticulitis, peptic ulceration, gastrointestinal motility or gastricacid secretion; dry mouth; mydriasis, tachycardia; ophthalmicinterventions cardiovascular disorders such as vagally induced sinusbradycardia.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES Example 1 Preparation of imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester

Under a nitrogen atmosphere, 7.0 g (55.0 mmol) of (R)-3-quinuclidinolwere suspended in 100 ml of CH₂Cl₂. After cooling the suspension to 0°C., a solution of 1,1′-carbonyldiimidazole (10.7 g, 66.0 mmol) in 150 mLof CH₂Cl₂ was added. The reaction mixture was stirred at 0° C. for 4hours, then water was added (100 ml). The organic layer was separated,washed with water, dried over sodium sulphate and the solvent wasevaporated under reduced pressure. The residue oil (13.1 g) was usedwithout further purification.

Example 2 Preparation of carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-ylester bis-(3-fluoro-phenyl)-methyl ester

A solution of 3,3′-difluorobenzhydrol (1.1 g, 5 mmol) in dry THF (8 mL)was added to a solution of BuLi (butyl lithium) 2.5M in hexane (2.0 mL,5 mmol) at 0° C.; after stirring for 30 minutes, a solution ofimidazole-1-carboxylic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester (1.1g, 5 mmol), obtained as described in example 1, in dry THF (16 ml) wasadded, and the mixture was stirred at 0° C. for 1.5 hours. Water (100ml) was added, and the mixture was extracted with EtOAc (2×50 ml). Thecollected organic layers were dried over sodium sulphate and the solventwas evaporated under reduced pressure. The residue was purified bycolumn chromatography on silica gel, eluting with CH₂Cl₂/MeOH=95/5.Fractions containing the product were combined and concentrated byevaporation to give 1.4 g, (75% yield) of the title compound as paleyellow oil.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.31 (m, 8H), 6.72 (s, 1H), 4.66 (m, 1H), 3.11 (m, 1H), 2.58 (m, 5H),1.96 (d, 1H, J=2.98 Hz), 1.59 (m, 4H).

The following intermediates (Examples 3 to 13) were obtained with aprocess similar to the one used to prepare the compound of Example 2, byreacting the compound prepared in Example 1 with the suitable alcohols,such as benzhydrol, fluorenol, 3-fluoro-benzol, 4-fluoro-benzol,4-bromo-benzol, 2-hydroxy biphenyl, 2-hydroxy methyl thiophene,bis-(4-chloro-phenol) or bis-(4-bromo-phenol).

Example 3 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbenzhydryl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with benzhydrol.

Example 4 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester9H-fluoren-9-yl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with fluorenol.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.86 (d, 2H, J=7.5 Hz), 7.63 (t, 2H, J=6.5 Hz), 7.48 (t, 2H, J=7.5Hz), 7.36 (m, 2H), 6.60 (s, 1H), 4.77 (t, 1H, J=4.2 Hz), 3.19 (dd, 1H,J=8.3, 14.9 Hz), 2.65 (m, 5H), 2.04 (d, 1H, J=2.9 Hz), 1.55 (m, 4H).

Example 5 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester3-fluoro-benzyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 3-fluoro-benzol.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.43 (m, 1H), 7.18 (m, 3H), 5.15 (s, 2H), 4.66 (m, 1H), 3.11 (m, 1H),2.54 (m, 4H), 1.96 (m, 1H), 1.49 (m, 5H).

Example 6 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester4-fluoro-benzyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 4-fluoro-benzol.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.46 (m, 2H), 7.22 (m, 2H), 5.11 (s, 2H), 4.63 (m, 1H), 2.61 (m, 5H),1.95 (d, 1H, J=3.0 Hz), 1.48 (m, 5H).

Example 7 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester4-bromo-benzyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 4-bromo-benzol.

Example 8 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbiphenyl-2-yl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 2-hydroxy biphenyl.

Example 9 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterthiophen-2-ylmethyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 2-hydroxy methyl thiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.62 (m, 1H), 7.24 (m, 1H), 7.07 (M, 1H), 5.33 (s, 2H), 4.67 (m, 1H),3.14 (m, 1H), 2.64 (m, 5H), 1.98 (d, 1H, J=3.1 Hz), 1.30 (m, 4H).

Example 10 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbis-(4-chloro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with bis-(4-chloro-phenol).

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.47 (m, 8H), 6.74 (s, 1H), 4.66 (t, 1H, J=4.0 Hz), 3.13 (m, 1H),2.66 (m, 5H), 1.97 (d, 1H, J=3.0 Hz), 1.51 (m, 4H).

Example 11 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbis-(4-bromo-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with bis-(4-bromo-phenol).

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 7.59 (m, 4H), 7.37 (m, 4H), 6.68 (s, 1H), 4.63 (t, 1H, J=3.0 Hz),3.30 (s, 1H), 3.08 (m, 1H), 2.64 (m, 4H), 1.94 (d, 1H, J=3.0 Hz), 1.59(m, 4H)

Example 12 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbis-(4-fluoro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with bis-(4-fluoro-phenol).

¹H NMR analysis (300 MHz, CDCl₃-d6)

δ: 7.34 (m, 4H), 7.06 (m, 4H), 6.67 (s, 1H), 4.72 (m, 1H), 3.23 (m, 1H),2.83 (m, 5H), 2.08 (m, 1H), 1.49 (m, 4H).

Example 13 Carbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl ester3-bromo-benzyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with 3-bromo-benzol.

Example 14 Preparation of(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 1)

The desired product was prepared by reacting 1.4 g, (3.7 mmol) ofcarbonic acid (R)-1-aza-bicyclo[2.2.2]oct-3-yl esterbis-(3-fluoro-phenyl)-methyl ester (obtained as described in example 2)with 2-(2-chloro)acetylthiophene (602 mg, 3.7 mmol) in EtOAc (5 ml).After stirring for 8 hours at room temperature, the solid precipitatedwas filtered, washed with Et₂O (2 ml) and dried under vacuum at 50° C.1.3 g of the title compound was obtained as white solid.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.06-1.97 (4H, m), 2.42 (1H, br s), 3.86-3.71 (6H, m), 4.25 (1H, m),5.13 (2H, s), 6.78 (1H, s), 7.47-7.15 (9H, m), 8.11 (1H, d), 8.22 (1H,d).

The following compounds were prepared using the route described inExample 14 and in particular by reacting the compounds described inExamples 3 to 13 with 2-(2-chloro)acetylthiophene, 1-phenethyl bromide,1-benzyl chloride, methyl iodide, or 1-(3-phenoxy-propyl) bromide, asdescribed in the following.

Example 15(R)-3-Benzhydryloxycarbonyloxy-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 2)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester benzhydryl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.03-1.95 (4H, m), 2.40 (1H, s), 3.88-3.65 (5H, m), 4.19 (1H, m),5.13 (1H, br s), 5.24 (2H, s), 6.74 (1H, s), 7.45-7.30 (11H, m), 8.16(1H, d), 8.22 (1H, d).

Example 16(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 3)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 9H-fluoren-9-yl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.09-1.98 (4H, m), 2.50 (1H, s), 3.78-3.69 (4H, m), 3.95 (1H, d),4.28 (1H, m), 5.27-5.22 (3H, br s), 6.52 (1H, s), 7.39-7.35 (3H, m),7.50 (2H, d), 7.67 (2H, t), 7.87 (2H, d), 8.17 (1H, d), 8.23 (1H, d).

Example 17(R)-3-(3-Fluoro-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 4)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 3-fluoro-benzyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.07-1.93 (4H, m), 2.42 (1H, s), 3.93-3.66 (5H, m), 4.27-4.21 (1H,m), 5.14 (1H, s), 5.22 (2H, s), 5.30 (2H, s), 7.50-7.18 (5H, m), 8.21(2H, m).

Example 18(R)-3-(4-Fluoro-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 5)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 4-fluoro-benzyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.09-1.89 (4H, m), 2.41 (1H, br s), 3.89-3.67 (5H, m), 4.25-4.18 (1H,m), 5.13 (1H, br s), 5.18 (2H, s), 5.25 (2H, s), 7.24 (2H, t), 7.35 (1H,t), 7.52-7.47 (2H, m), 8.17 (1H, d), 8.22 (1H, d).

Example 19(R)-3-(4-Bromo-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 6)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 4-bromo-benzyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.06-1.93 (4H, m), 2.41 (1H, s), 3.91-3.35 (5H, m), 4.26-4.19 (1H,m), 5.12 (1H, s), 5.17 (2H, s), 5.27 (2H, s), 7.41-7.34 (3H, m), 7.61(2H, d), 8.17 (1H, d), 8.22 (1H, d).

Example 20(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 7)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester biphenyl-2-yl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.03-1.88 (4H, m), 2.23 (1H, s), 3.79-3.60 (5H, m), 4.17-4.10 (1H,m), 5.09 (1H, br s), 5.16 (2H, s), 7.49-7.36 (10H, m), 8.14 (1H, d),8.23 (1H, d).

Example 21(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-phenethyl-1-azonia-bicyclo[2.2.2]octane;bromide (compound 8)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 9H-fluoren-9-yl ester with1-phenethyl bromide.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.04-1.92 (4H, m), 2.50 (2H, s), 3.09-3.00 (2H, m), 3.70-3.43 (7H,m), 4.06-3.99 (1H, m), 5.22 (1H, s), 6.65 (1H, s), 7.89-7.26 (13H, m).

Example 22(R)-1-Benzyl-3-(9H-fluoren-9-yloxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 9)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 9H-fluoren-9-yl ester with1-benzyl chloride.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 1.99-1.81 (4H, m), 2.42 (1H, s), 3.51-3.41 (4H, m), 3.71 (1H, m),3.92 (1H, m), 4.63 (2H, q), 5.16 (1H, s), 6.64 (1H, s), 7.68-7.36 (11H,m), 7.87 (2H, d).

Example 23(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-methyl-1-azonia-bicyclo[2.2.2]octane;iodide (compound 10)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 9H-fluoren-9-yl ester with methyliodide.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 1.99 (5H, m), 2.50 (1H, s), 3.00 (3H, s), 3.44 (3H, m), 3.47 (1H, d),3.67 (1H, m), 5.25 (1H, m), 6.64 (1H, s), 7.88-7.36 (8H, m).

Example 24(R)-3-(9H-Fluoren-9-yloxycarbonyloxy)-1-(3-phenoxy-propyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 11)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 9H-fluoren-9-yl ester with1-(3-phenoxy-propyl) bromide.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.19-1.90 (7H, m), 2.50 (1H, s), 3.68-3.32 (6H, m), 4.05-3.96 (3H,m), 5.18 (1H, s), 6.62 (1H, s), 7.88-6.94 (13H, m).

Example 25(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-(3-phenoxy-propyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 12)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester biphenyl-2-yl ester with1-(3-phenoxy-propyl) bromide.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.20-1.83 (7H, m), 2.50 (2H, s), 3.55-3.28 (5H, m), 3.90-3.83 (1H,m), 4.05 (2H, t), 4.98 (1H, br s), 6.99-6.95 (3H, m), 7.50-7.30 (11H,m).

Example 26(R)-3-(Biphenyl-2-yloxycarbonyloxy)-1-methyl-1-azonia-bicyclo[2.2.2]octane;iodide (compound 13)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester biphenyl-2-yl ester with methyliodide.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.18-1.80 (6H, m), 2.95 (3H, s), 3.47-3.32 (4H, m), 3.84-3.75 (1H,m), 4.98-4.96 (1H, m), 5.52-7.39 (9H, m).

Example 27(R)-1-(2-oxo-2-thiophen-2-yl-ethyl)-3-(thiophen-2-ylmethoxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 14)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester thiophen-2-ylmethyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.10-1.95 (4H, m), 2.44 (1H, br s), 3.87-3.66 (5H, m), 4.25-4.16 (1H,m), 5.16 (3H, s), 5.40 (2H, s), 7.66-7.08 (4H, m), 8.15 (1H, dd), 8.24(1H, dd).

Example 28(R)-3-[Bis-(4-chloro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 15)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(4-chloro-phenyl)-methylester with 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 1.90-2.06 (4H, m), 2.41 (1H, s), 3.61-3.85 (5H, m), 4.14 (1H, s),5.12 (3H, s), 7.35 (1H, t), 7.47 (9H, d), 8.11 (1H, d), 8.22 (1H, d).

Example 29(R)-3-[Bis-(4-bromo-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 16)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(4-bromo-phenyl)-methyl esterwith 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.09-1.96 (4H, m), 2.40 (1H, s), 3.78-3.72 (5H, m), 3.86-3.82 (1H,m), 5.16-5.13 (3H, m), 6.75 (1H, s), 7.43-7.35 (5H, m), 7.63-7.59 (4H,m), 8.12 (1H, d), 8.22 (1H, d).

Example 30(R)-3-[Bis-(4-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 17)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(4-fluoro-phenyl)-methylester with 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.09-1.96 (4H, m), 2.40 (1H, br s), 3.89-3.65 (5H, m), 4.23-4.16 (1H,m), 5.14 (1H, d), 5.23 (2H, d), 6.79 (1H, s), 7.52-7.20 (9H, m), 8.16(1H, d), 8.22 (1H, d).

Example 31(R)-3-(3-Bromo-benzyloxycarbonyloxy)-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 18)

The desired product was prepared by reacting carbonic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester 3-bromo-benzyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d6)

δ: 2.14-1.96 (4H, m), 2.45 (1H, br s), 3.91-3.64 (5H, m), 4.26-4.19 (1H,m), 5.23-5.16 (5H, m), 7.59-7.37 (3H, m), 7.67-7.60 (2H, m), 8.17 (1H,d), 8.25 (1H, d).

Example 32 Preparation of carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(2-fluoro-phenyl)-methyl ester

NaH (60% dispersion in mineral oil, 36.3 mg, 0.9 mmol) was added to asolution of (2-fluoro-phenyl)-(3-fluoro-phenyl)-methanol (200 mg, 0.9mmol) in anhydrous THF (3 ml), at 0° C. The reaction mixture was stirredat this temperature for 30 minutes, then (R)-quinuclidin-3-yl1H-imidazole-1-carboxylate (201 mg, 0.9 mmol), obtained as described inExample 1, was added and the stirring continued at room temperature for1 hour. Then a saturated solution of NH₄Cl was added, and the aqueousphase was extracted with EtOAc (3×30 ml). The organic phase was driedover anhydrous sodium sulfate, and the solvent was evaporated underreduced pressure. The resulting crude was purified by flashchromatography on silica gel eluting with EtOAc/MeOH=9/1. The desiredproduct was collected as colorless viscous oil (244 mg; 72% yield;mixture of diasteroisomers).

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.34-7.57 (m, 3H), 7.11-7.33 (m, 5H), 6.87 (s, 1H), 4.50-4.81 (m,1H), 3.13 (dd, 1H), 2.53-2.81 (m, 5H), 1.91-2.08 (m, 1H), 1.54-1.76 (m,2H), 1.41-1.54 (m, 1H), 1.26-1.41 (m, 1H).

LC-MS (ESI POS): 374.14 (MH+).

The following intermediates (example 33 to 42) were obtained with aprocess similar to the one used to prepare the compound of Example 32,by reacting the compound prepared in Example 1 with the suitablealcohols, such as bis(2-chlorophenyl)methanol, phenyl-o-tolyl-methanol,(3-fluoro-phenyl)-(3-methoxy-phenyl)-methanol,cyclohexyl-(3-fluoro-phenyl)-methanol,(3-chloro-phenyl)-(3-fluoro-phenyl)-methanol,(3,5-difluoro-phenyl)-(3-fluoro-phenyl)-methanol,(3-fluoro-phenyl)-m-tolyl-methanol,(3-fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methanol,(3-fluoro-phenyl)-(4-fluoro-phenyl)-methanol,(3,4-difluoro-phenyl)-phenyl-methanol, and(3-fluoro-phenyl)-phenyl-methanol.

Example 33 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) esterbis-(2-chloro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with bis(2-chlorophenyl)methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.51-7.63 (m, 2H), 7.37-7.51 (m, 4H), 7.22-7.31 (m, 2H), 7.21 (s,1H), 4.53-4.77 (m, 1H), 3.13 (dd, 1H), 2.54-2.81 (m, 5H), 1.88-2.04 (m,1H), 1.54-1.78 (m, 2H), 1.41-1.54 (m, 1H), 1.27-1.41 (m, 1H).

LC-MS (ESI POS): 406.05 (MH+).

Example 34 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) esterphenyl-o-tolyl-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with phenyl-o-tolyl-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.09-7.51 (m, 9H), 6.80 (s, 1H), 4.47-4.79 (m, 1H), 3.10 (dd, 1H),2.54-2.77 (m, 5H), 2.27 (s, 3H), 1.94 (br. s., 1H), 1.52-1.76 (m, 2H),1.38-1.52 (m, 1H), 1.16-1.38 (m, 1H).

LC-MS (ESI POS): 352.13 (MH+).

Example 35 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(3-methoxy-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-fluoro-phenyl)-(3-methoxy-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.37-7.51 (m, 1H), 7.21-7.35 (m, 3H), 7.08-7.20 (m, 1H), 6.94-7.04(m, 2H), 6.82-6.92 (m, 1H), 6.65 (s, 1H), 4.53-4.79 (m, 1H), 3.75 (s,3H), 3.11 (dd, 1H), 2.54-2.79 (m, 5H), 1.86-2.01 (m, 1H), 1.53-1.76 (m,2H), 1.40-1.53 (m, 1H), 1.21-1.40 (m, 1H).

LC-MS (ESI POS): 386.05 (MH+).

Example 36 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) estercyclohexyl-(3-fluoro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester withcyclohexyl-(3-fluoro-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.33-7.49 (m, 1H), 7.02-7.21 (m, 3H), 5.31 (d, 1H), 4.34-4.69 (m,1H), 2.99-3.21 (m, 1H), 2.43-2.79 (m, 4H), 1.85-1.98 (m, 1H), 0.83-1.83(m, 16H).

LC-MS (ESI POS): 362.16 (MH+).

Example 37 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-chloro-phenyl)-(3-fluoro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-chloro-phenyl)-(3-fluoro-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.50-7.56 (m, 1H) 7.35-7.49 (m, 4H) 7.25-7.35 (m, 2H) 7.08-7.21 (m,1H) 6.71 (s, 1H) 4.58-4.71 (m, 1H) 3.02-3.17 (m, 1H) 2.54-2.78 (m, 4H)1.91-1.98 (m, 1H) 1.40-1.70 (m, 3H) 1.21-1.38 (m, 2H).

LC-MS (ESI POS): 390.04 (MH+).

Example 38 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3,5-difluoro-phenyl)-(3-fluoro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3,5-difluoro-phenyl)-(3-fluoro-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.40-7.60 (m, 2H) 7.10-7.26 (m, 5H) 6.97 (d, 1H) 4.58-4.73 (m, 1H)3.01-3.19 (m, 1H) 2.54-2.72 (m, 5H) 1.85-2.06 (m, 1H) 1.40-1.69 (m, 3H)1.23-1.38 (m, 1H).

LC-MS (ESI POS): 392.05 (MH+).

Example 39 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-m-tolyl-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-fluoro-phenyl)-m-tolyl-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.36-7.49 (m, 1H), 7.06-7.32 (m, 7H), 6.64 (s, 1H), 4.32-4.80 (m,1H), 3.10 (dd, 1H), 2.54-2.77 (m, 5H), 2.30 (s, 3H), 1.87-2.04 (m, 1H),1.11-1.76 (m, 4H).

LC-MS (ESI POS): 370.1 (MH+).

Example 40 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.31-7.49 (m, 3H), 7.20-7.30 (m, 4H), 7.07-7.20 (m, 1H), 6.66 (s,1H), 4.64 (ddd, 1H), 3.10 (dd, 1H), 2.54-2.76 (m, 5H), 2.46 (s, 3H),1.86-2.02 (m, 1H), 1.38-1.77 (m, 3H), 1.16-1.38 (m, 1H).

LC-MS (ESI POS): 401.93 (MH+).

Example 41 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(4-fluoro-phenyl)-(3-fluoro-phenyl)-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-fluoro-phenyl)-(4-fluoro-phenyl)-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.36-7.56 (m, 3H), 7.03-7.32 (m, 5H), 6.72 (s, 1H), 4.64 (ddd, 1H),3.10 (dd, 1H), 2.65-2.80 (m, 2H), 2.54-2.65 (m, 3H), 1.95 (br. s., 1H),1.53-1.74 (m, 2H), 1.40-1.53 (m, 1H), 1.18-1.40 (m, 1H).

LC-MS (ESI POS): 374.14 (MH+).

Example 42 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3,4-difluoro-phenyl)-phenyl-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3,4-difluoro-phenyl)-phenyl-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.47-7.62 (m, 1H), 7.20-7.47 (m, 7H), 6.69 (s, 1H), 4.64 (m, 1H),3.10 (dd, 1H), 2.54-2.79 (m, 5H), 1.87-2.01 (m, 1H), 1.53-1.74 (m, 2H),1.39-1.53 (m, 1H), 1.17-1.39 (m, 1H).

LC-MS (ESI POS): 373.96 (MH+).

Example 43 Carbonic acid (R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-phenyl-methyl ester

The desired product was prepared by reacting imidazole-1-carboxylic acid(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester with(3-fluoro-phenyl)-phenyl-methanol.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.21-7.55 (m, 7H), 7.00-7.20 (m, 2H), 6.69 (s, 1H), 4.52-4.73 (m,1H), 3.10 (dd, 1H), 2.54-2.78 (m, 5H), 1.89-1.97 (m, 1H), 1.52-1.75 (m,2H), 1.40-1.52 (m, 1H), 1.19-1.39 (m, 1H).

LC-MS (ESI POS): 356.16 (MH+).

The following compounds were prepared using the route described inExample 14 and in particular by reacting the compound described inExample 2 with 2-bromo-1-phenyl-ethanone, 2-bromo-acetamide,2-bromo-1-p-tolyl-ethanone, 2-bromo-1-(4-fluoro-phenyl)-ethanone,2-bromo-1-(4-methoxy-phenyl)-ethanone, 2-bromo-1-thiophen-3-yl-ethanone,2-bromo-N-phenyl-acetamide, 2-bromo-1-(5-chloro-thiophen-2-yl)-ethanone,2-bromo-1-(3,5-dibromo-thiophen-2-yl)-ethanone,2-bromo-1-thiazol-2-yl-ethanone, 2-bromo-1-o-tolyl-ethanone,2-bromo-1-m-tolyl-ethanone, 5-bromo-2-methylpent-2-ene,1-benzo[b]thiophen-2-yl-2-bromo-ethanone, bromomethyl-benzene,(2-bromoethoxy)benzene, or methyl iodide, as described in the following.

Example 44(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-phenyl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 19)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-phenyl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.89-8.06 (m, 2H), 7.70-7.86 (m, 1H), 7.56-7.70 (m, 2H), 7.39-7.55(m, 2H), 7.26-7.39 (m, 4H), 7.07-7.26 (m, 2H), 6.79 (s, 1H), 5.19 (s,2H), 5.13-5.18 (m, 1H), 4.03-4.27 (m, 1H), 3.56-3.86 (m, 5H), 2.35-2.47(m, 1H), 1.91-2.16 (m, 4H).

LC-MS (ESI POS): 492.10 (MH+).

Example 45(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-carbamoylmethyl-1-azonia-bicyclo[2.2.2]octane;bromide (compound 20)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-acetamide.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.92 (br. s., 1H), 7.69 (br. s., 1H), 7.38-7.53 (m, 2H), 7.25-7.38(m, 4H), 7.06-7.25 (m, 2H), 6.77 (s, 1H), 5.10 (br. s., 1H), 4.02-4.20(m, 1H), 3.99 (s, 2H), 3.76-3.85 (m, 1H), 3.49-3.74 (m, 4H), 2.32-2.43(m, 1H), 1.83-2.09 (m, 4H).

LC-MS (ESI POS): 431.10 (MH+).

Example 46(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-p-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 21)

The desired product is prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-p-tolyl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.73-8.03 (m, 2H), 7.26-7.50 (m, 8H), 7.03-7.25 (m, 2H), 6.78 (s,1H), 5.15-5.21 (m, 1H), 5.14 (s, 2H), 4.07-4.26 (m, 1H), 3.76-3.93 (m,1H), 3.48-3.76 (m, 4H), 2.43-2.47 (m, 1H), 2.42 (s, 3H), 1.91-2.15 (m,4H).

LC-MS (ESI POS): 505.95 (MH+).

Example 47(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(4-fluoro-phenyl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane;bromide (compound 22)

The desired product is prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-(4-fluoro-phenyl)-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.98-8.18 (m, 2H), 7.40-7.58 (m, 4H), 7.25-7.40 (m, 4H), 7.05-7.25(m, 2H), 6.78 (s, 1H), 5.17 (s, 2H), 5.13-5.24 (m, 1H), 4.08-4.27 (m,1H), 3.55-3.86 (m, 5H), 2.34-2.47 (m, 1H), 1.90-2.17 (m, 4H).

LC-MS (ESI POS): 509.94 (MH+).

Example 48(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(4-methoxy-phenyl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane;bromide (compound 23)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-(4-methoxy-phenyl)-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.92-8.03 (m, 2H), 7.25-7.52 (m, 6H), 6.95-7.25 (m, 4H), 6.78 (s,1H), 5.13-5.24 (m, 1H), 5.10 (s, 2H), 4.05-4.27 (m, 1H), 3.88 (s, 3H),3.48-3.87 (m, 5H), 2.33-2.46 (m, 1H), 1.90-2.15 (m, 4H).

LC-MS (ESI POS): 521.98 (MH+).

Example 49(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-3-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 24)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-thiophen-3-yl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.62 (dd, 1H), 7.74 (dd, 1H), 7.56 (dd, 1H), 7.39-7.52 (m, 2H),7.27-7.39 (m, 4H), 7.09-7.25 (m, 2H), 6.78 (s, 1H), 5.10-5.27 (m, 1H),5.03 (s, 2H), 4.14 (dd, 1H), 3.45-3.94 (m, 5H), 2.36-2.47 (m, 1H),1.70-2.19 (m, 4H).

LC-MS (ESI POS): 497.89 (MH+).

Example 50(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-phenylcarbamoylmethyl-1-azonia-bicyclo[2.2.2]octane;bromide (compound 25)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-N-phenyl-acetamide.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 10.52 (s, 1H), 7.52-7.63 (m, 2H), 7.26-7.51 (m, 8H), 7.01-7.23 (m,3H), 6.78 (s, 1H), 4.91-5.31 (m, 1H), 4.26 (d, 1H), 4.19 (d, 1H),4.02-4.20 (m, 1H), 3.81-3.95 (m, 1H), 3.51-3.81 (m, 4H), 2.43 (dd, 1H),1.71-2.17 (m, 4H).

LC-MS (ESI POS): 506.94 (MH+).

Example 51(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(5-chloro-thiophen-2-yl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane;bromide (compound 26)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-(5-chloro-thiophen-2-yl)-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.00 (d, 1H), 7.38-7.60 (m, 3H), 7.25-7.38 (m, 4H), 7.02-7.25 (m,2H), 6.78 (s, 1H), 5.06-5.36 (m, 1H), 4.99 (s, 2H), 3.97-4.24 (m, 1H),3.51-3.84 (m, 5H), 2.36-2.45 (m, 1H), 1.89-2.15 (m, 4H).

LC-MS (ESI POS): 531.82 (MH+).

Example 52(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-[2-(3,5-dibromo-thiophen-2-yl)-2-oxo-ethyl]-1-azonia-bicyclo[2.2.2]octane;chloride (compound 27)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-(3,5-dibromo-thiophen-2-yl)-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.68 (s, 1H), 7.40-7.52 (m, 2H), 7.26-7.39 (m, 4H), 7.11-7.23 (m,2H), 6.78 (s, 1H), 5.11-5.23 (m, 1H), 5.08 (s, 2H), 4.07-4.32 (m, 1H),3.49-4.00 (m, 5H), 2.35-2.46 (m, 1H), 1.74-2.19 (m, 4H).

LC-MS (ESI POS): 653.62 (MH+).

Example 53(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiazol-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 28)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-thiazol-2-yl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.39 (d, 1H), 8.24 (d, 1H), 7.39-7.52 (m, 2H), 7.26-7.39 (m, 4H),7.10-7.25 (m, 2H), 6.78 (s, 1H), 5.22 (s, 2H), 4.98-5.19 (m, 1H),4.07-4.33 (m, 1H), 3.52-4.01 (m, 5H), 2.36-2.45 (m, 1H), 1.80-2.17 (m,4H).

LC-MS (ESI POS): 499.04 (MH+).

Example 54(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-o-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 29)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-o-tolyl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.66-7.92 (m, 1H), 7.52-7.65 (m, 1H), 7.26-7.51 (m, 8H), 7.09-7.24(m, 2H), 6.79 (s, 1H), 5.11-5.24 (m, 1H), 5.08 (s, 2H), 4.06-4.26 (m,1H), 3.53-3.92 (m, 5H), 2.46 (s, 3H), 2.36-2.45 (m, 1H), 1.83-2.19 (m,4H).

LC-MS (ESI POS): 505.95 (MH+).

Example 55(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-m-tolyl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 30)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 2-bromo-1-m-tolyl-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.70-7.84 (m, 2H), 7.39-7.64 (m, 4H), 7.26-7.39 (m, 4H), 7.10-7.24(m, 2H), 6.79 (s, 1H), 5.17 (s, 2H), 5.15-5.26 (m, 1H), 4.00-4.29 (m,1H), 3.48-3.90 (m, 5H), 2.42-2.46 (m, 1H), 2.41 (s, 3H), 1.86-2.15 (m,4H).

LC-MS (ESI POS): 506.11 (MH+).

Example 56(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(4-methyl-pent-3-enyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 31)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 5-bromo-2-methylpent-2-ene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.39-7.52 (m, 2H), 7.25-7.39 (m, 4H), 7.08-7.23 (m, 2H), 6.77 (s,1H), 5.03-5.09 (m, 1H), 4.89-5.02 (m, 1H), 3.74-3.99 (m, 1H), 3.48-3.60(m, 1H), 3.31-3.47 (m, 4H), 3.15 (t, 2H), 2.29-2.43 (m, 3H), 1.77-2.13(m, 4H), 1.68 (s, 3H), 1.63 (s, 3H).

LC-MS (ESI POS): 456.00 (MH+).

Example 57(R)-1-(2-Benzo[b]thiophen-2-yl-2-oxo-ethyl)-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-azonia-bicyclo[2.2.2]octane;bromide (compound 32)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with 1-benzo[b]thiophen-2-yl-2-bromo-ethanone.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.50 (s, 1H), 8.01-8.26 (m, 2H), 7.58-7.67 (m, 1H), 7.51-7.58 (m,1H), 7.40-7.51 (m, 2H), 7.26-7.40 (m, 4H), 7.08-7.24 (m, 2H), 6.79 (s,1H), 5.19 (s, 2H), 5.10-5.17 (m, 1H), 4.09-4.26 (m, 1H), 3.81-3.93 (m,1H), 3.57-3.81 (m, 4H), 2.32-2.47 (m, 1H), 1.84-2.19 (m, 4H).

LC-MS (ESI POS): 547.92 (MH+).

Example 58(R)-1-Benzyl-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-azonia-bicyclo[2.2.2]octane;bromide (compound 33)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with bromomethyl-benzene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.39-7.60 (m, 7H), 7.24-7.37 (m, 4H), 7.11-7.24 (m, 2H), 6.77 (s,1H), 4.86-5.18 (m, 1H), 4.51 (d, 1H), 4.45 (d, 1H), 3.68-3.84 (m, 1H),3.52-3.65 (m, 1H), 3.41-3.52 (m, 2H), 3.32-3.38 (m, 2H), 2.30-2.41 (m,1H), 1.63-2.16 (m, 4H).

LC-MS (ESI POS): 463.98 (MH+).

Example 59(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-phenoxy-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 34)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with (2-bromoethoxy)benzene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.39-7.51 (m, 2H), 7.26-7.37 (m, 6H), 7.11-7.23 (m, 2H), 6.94-7.06(m, 3H), 6.78 (s, 1H), 4.92-5.20 (m, 1H), 4.44 (br. s., 2H), 3.92-4.10(m, 1H), 3.67-3.75 (m, 2H), 3.41-3.67 (m, 5H), 2.33-2.42 (m, 1H),1.74-2.15 (m, 4H).

LC-MS (ESI POS): 493.97 (MH+).

Example 60(R)-3-[Bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-methyl-1-azonia-bicyclo[2.2.2]octane;iodide (compound 35)

The desired product was prepared by reacting(R)-1-aza-bicyclo[2.2.2]oct-3-yl ester bis-(3-fluoro-phenyl)-methylester with methyl iodide.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 7.40-7.52 (m, 2H), 7.25-7.37 (m, 4H), 7.10-7.25 (m, 2H), 6.77 (s,1H), 4.87-5.16 (m, 1H), 3.84 (dd, 1H), 3.57 (dt, 1H), 3.32-3.50 (m, 4H),2.96 (s, 3H), 2.32-2.40 (m, 1H), 1.63-2.21 (m, 4H).

LC-MS (ESI POS): 387.97 (MH+).

Example 61 Preparation of(R)-3-[bis-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-pyridin-4-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;bromide (compound 36)

N,N-Diisopropylethylamine (0.037 mL, 0.22 mmol) was added to a solutionof 2-bromo-1-(pyridin-4-yl)ethanone hydrobromide (60.2 mg, 0.22 mmol)dissolved in EtOAc (0.5 mL). After 10 minutes, a solution of(R)-bis(3-fluorophenyl)methyl quinuclidin-3-yl carbonate (80 mg, 0.22mmol), obtained as described in Example 2, in EtOAc (0.5 mL) was added,and the reaction mixture was stirred at room temperature for 2 hours.Et₂O (1 mL) was added, and the precipitate was collected by suctionfiltration. The solid was dissolved in water and extracted with DCM. Theorganic phase was dried over anhydrous sodium sulfate and filtered, andthe solvent was evaporated under reduced pressure to give the desiredcompound as a pink solid (76 mg; 62% yield).

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.47-9.13 (m, 2H), 7.67-7.95 (m, 2H), 7.39-7.56 (m, 2H), 7.25-7.39(m, 4H), 7.06-7.25 (m, 2H), 6.78 (s, 1H), 5.19 (s, 2H), 5.07-5.18 (m,1H), 4.00-4.30 (m, 1H), 3.47-3.89 (m, 5H), 2.39-2.46 (m, 1H), 1.88-2.22(m, 4H).

LC-MS (ESI POS): 492.97 (MH+).

Example 62(R)-3-[(2-Fluoro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 37)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(2-fluoro-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (d, 1H), 7.97-8.15 (m, 1H), 7.10-7.59 (m, 9H), 6.93 (s, 1H),5.12-5.27 (m, 1H), 5.07 (s, 2H), 4.05-4.28 (m, 1H), 3.52-3.95 (m, 5H),2.38-2.46 (m, 1H), 1.71-2.18 (m, 4H).

LC-MS (ESI POS): 498.02 (MH+).

Example 63(R)-3-[Bis-(2-chloro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 38)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester bis-(2-chloro-phenyl)-methylester with 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.09 (dd, 1H), 7.52-7.69 (m, 2H), 7.39-7.53 (m, 4H),7.27-7.39 (m, 3H), 7.26 (s, 1H), 5.12-5.29 (m, 1H), 5.09 (s, 2H), 4.15(dd, 1H), 3.45-3.95 (m, 5H), 2.45 (br. s., 1H), 1.86-2.17 (m, 4H).

LC-MS (ESI POS): 529.96 (MH+).

Example 64(R)-1-(2-oxo-2-thiophen-2-yl-ethyl)-3-(phenyl-o-tolyl-methoxycarbonyloxy)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 39)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester phenyl-o-tolyl-methyl esterwith 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (d, 1H), 7.99-8.14 (m, 1H), 7.12-7.53 (m, 10H), 6.85 (s, 1H),5.09-5.21 (m, 1H), 5.07 (s, 2H), 3.94-4.23 (m, 1H), 3.44-3.90 (m, 5H),2.34-2.47 (m, 1H), 2.29 (s, 3H), 1.77-2.16 (m, 4H).

LC-MS (ESI POS): 476.08 (MH+).

Example 65(R)-3-[(3-Fluoro-phenyl)-(3-methoxy-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 40)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(3-methoxy-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.09 (d, 1H), 7.38-7.51 (m, 1H), 7.24-7.38 (m, 4H),7.09-7.22 (m, 1H), 6.97-7.06 (m, 2H), 6.84-6.95 (m, 1H), 6.72 (s, 1H),5.12-5.20 (m, 1H), 5.09 (s, 2H), 4.06-4.22 (m, 1H), 3.78-3.89 (m, 1H),3.76 and 3.75 (s, 3H), 3.52-3.73 (m, 4H), 2.36-2.46 (m, 1H), 1.84-2.18(m, 4H).

LC-MS (ESI POS): 510.08 (MH+).

Example 66(R)-3-[Cyclohexyl-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 41)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) estercyclohexyl-(3-fluoro-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.12-8.31 (m, 1H), 7.88-8.15 (m, 1H), 7.29-7.53 (m, 2H), 6.96-7.24(m, 3H), 5.30-5.47 (m, 1H), 5.02-5.20 (m, 3H), 3.99-4.25 (m, 1H),3.47-3.89 (m, 5H), 1.45-2.21 (m, 9H), 0.79-1.45 (m, 7H).

LC-MS (ESI POS): 486.11 (MH+).

Example 67(R)-3-[(3-Chloro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 42)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-chloro-phenyl)-(3-fluoro-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.08 (dd, 1H), 7.52-7.61 (m, 1H), 7.26-7.51 (m, 7H),7.11-7.23 (m, 1H), 6.78 (s, 1H), 5.11-5.20 (m, 1H), 5.06 (s, 2H),4.04-4.23 (m, 1H), 3.78-3.89 (m, 1H), 3.51-3.78 (m, 4H), 2.37-2.46 (m,1H), 1.69-2.21 (m, 4H).

LC-MS (ESI POS): 514.05 (MH+).

Example 68(R)-3-[(3,5-Difluoro-phenyl)-(3-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 43)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3,5-difluoro-phenyl)-(3-fluoro-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.08 (dd, 1H), 7.39-7.60 (m, 2H), 7.31-7.39 (m, 1H),7.12-7.29 (m, 5H), 7.02 (s, 1H), 5.11-5.26 (m, 1H), 5.06 (s, 2H),4.04-4.27 (m, 1H), 3.48-3.96 (m, 5H), 2.33-2.45 (m, 1H), 1.83-2.22 (m,4H).

LC-MS (ESI POS): 515.8 (MH+).

Example 69(R)-3-[(3-Fluoro-phenyl)-m-tolyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 44)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-m-tolyl-methyl ester with 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (d, 1H), 8.08 (d, 1H), 7.39-7.52 (m, 1H), 7.04-7.38 (m, 8H),6.71 (s, 1H), 5.13 (m, 1H), 5.06 (s, 2H), 4.02-4.23 (m, 1H), 3.50-3.92(m, 5H), 2.39-2.46 (m, 1H), 2.31 (s, 3H), 1.76-2.18 (m, 4H).

LC-MS (ESI POS): 494.05 (MH+).

Example 70(R)-3-[(3-Fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 45)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3-fluoro-phenyl)-(4-methylsulfanyl-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.09 (d, 1H), 7.32-7.59 (m, 4H), 7.22-7.32 (m, 4H),7.09-7.22 (m, 1H), 6.73 (s, 1H), 5.11-5.19 (m, 1H), 5.09 (s, 2H),4.05-4.23 (m, 1H), 3.51-3.89 (m, 5H), 2.46 (s, 3H), 2.37-2.44 (m, 1H),1.79-2.14 (m, 4H).

LC-MS (ESI POS): 525.91 (MH+).

Example 71(R)-3-[(3-Fluoro-phenyl)-(4-fluoro-phenyl)-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 46)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(4-fluoro-phenyl)-(3-fluoro-phenyl)-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.08 (dd, 1H), 7.40-7.58 (m, 3H), 7.35 (dd, 1H),7.08-7.33 (m, 5H), 6.78 (s, 1H), 5.10-5.19 (m, 1H), 5.05 (s, 2H),3.97-4.23 (m, 1H), 3.50-3.89 (m, 5H), 2.33-2.46 (m, 1H), 1.88-2.17 (m,4H).

LC-MS (ESI POS): 497.91 (MH+).

Example 72(R)-3-[(3,4-Difluoro-phenyl)-phenyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 47)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester(3,4-difluoro-phenyl)-phenyl-methyl ester with2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (ddd, 1H), 8.01-8.13 (m, 1H), 7.49-7.64 (m, 1H), 7.21-7.49 (m,8H), 6.76 (s, 1H), 5.11-5.21 (m, 1H), 5.09 (s, 2H), 4.02-4.28 (m, 1H),3.46-3.92 (m, 5H), 2.33-2.46 (m, 1H), 1.69-2.17 (m, 4H).

LC-MS (ESI POS): 497.90 (MH+).

Example 73(R)-3-[(3-Fluoro-phenyl)-phenyl-methoxycarbonyloxy]-1-(2-oxo-2-thiophen-2-yl-ethyl)-1-azonia-bicyclo[2.2.2]octane;chloride (compound 48)

The desired product was prepared by reacting carbonic acid(R)-(1-aza-bicyclo[2.2.2]oct-3-yl) ester (3-fluoro-phenyl)-phenyl-methylester with 2-(2-chloro)acetylthiophene.

¹H NMR analysis (300 MHz, DMSO-d₆)

δ: 8.21 (dd, 1H), 8.08 (ddd, 1H), 7.22-7.54 (m, 9H), 7.00-7.23 (m, 1H),6.76 (s, 1H), 5.11-5.22 (m, 1H), 5.06 (s, 2H), 4.01-4.22 (m, 1H),3.47-3.92 (m, 5H), 2.36-2.45 (m, 1H), 1.70-2.17 (m, 4H).

LC-MS (ESI POS): 479.94 (MH+).

Legend

*NMR

s=singlet

d=doublet

t=triplet

q=quartet

dd=doublet of doublets

m=multiplet

br=broad

Biological Characterization.

The interaction with M3 muscarinic receptors can be estimated by theresults of in vitro studies which evaluated the M3/M2 binding assays,the potency of the test compounds, and the offset of the inhibitoryactivity produced after washout of the antagonists in isolated guineapig trachea and by the in vivo duration of action againstacetylcholine-induced bronchospasm in the guinea pig.

Example 74 M3/M2 Binding Assays

CHO-K1 clone cells expressing the human M2 or M3-receptors (SwissprotP08172, P20309 respectively) were harvested in Ca⁺⁺/Mg⁺⁺ freephosphate-buffered saline and collected by centrifugation at 1500 rpmfor 3 minutes. The pellets were re-suspended in ice cold buffer A (15 mMTris-HCl pH 7.4, 2 mM MgCl₂, 0.3 mM EDTA, 1 mM EGTA) and homogenized bya PBI politron (setting 5 for 15 s). The crude membrane fraction wascollected by two consecutive centrifugation steps at 40000 g for 20minutes at 4° C., and separated by a washing step in buffer A. Thepellets obtained were finally resuspended in buffer B (75 mM Tris HCl pH7.4, 12.5 mM MgCl₂, 0.3 mM EDTA, 1 mM EGTA, 250 mM sucrose), andaliquots were stored at −80° C.

The day of experiment, frozen membranes were re-suspended in buffer C(50 mM Tris-HCl pH 7.4, 2.5 mM MgCl₂, 1 mM EDTA). The non-selectivemuscarinic radioligand [³H]-N-methyl scopolamine (Mol. Pharmacol.45:899-907) was used to label the M2, and M3 binding sites. Bindingexperiments were performed in duplicate (ten point concentrationscurves) in 96 well plates at radioligand concentration of 0.1-0.3 nM.The non-specific binding was determined in the presence of cold N-methylscopolamine 10 μM. Samples (final volume 0.75 ml) were incubated at roomtemperature for 60 minutes for M2 and 90 minutes for M3 binding assay.The reaction was terminated by rapid filtration through GF/B Unifilterplates and two washes (0.75 ml) with cold buffer C using a PackardFiltermate Harvester. Radioactivity on the filters was measured by amicroplate scintillation counter TriCarb 2500 (PerkinElmer).

Example 75 In Vitro Interaction with the M3 Receptors

The potency of the antagonist activity in isolated guinea pig tracheawas investigated following a method previously described by Haddad, E.B., et al. in Br. J. Pharmacol., vol. 127, pp. 413-420 (1999), with fewmodifications. A cumulative concentration-response curve to testantagonists was constructed on preparations pre-contracted by carbachol,until a complete inhibition of smooth muscle tone was achieved. Theconcentration of antagonist producing a 50% reversal ofcarbachol-induced tonic contraction (IC₅₀) was taken as a measure of itspotency in this bioassay.

In the experiments aiming at assessing the offset of the inhibitoryeffects produced by test compounds, the minimal concentration of thetest compounds known to produce a maximal inhibitory effect wasadministered to carbachol-pre-contracted preparations. As soon as thetonic contraction was completely reversed, the organ bath solution wasrenewed and preparations were thoroughly washed with fresh Krebssolution. Carbachol (0.3 μM) was administered again (at 30 minuteinterval between washout and next administration) during the next 4hours.

After the administration of carbachol, the inhibitory effects of thecompounds of the invention, administered at a concentration of 10 nM,were expressed as percentage of the recovery of the contracting responseto carbachol. The percentage of recovery four hours after the washoutwas lower than 50%.

The values of inhibitory M3 activity tested on compounds 1, 2, 3, 4, 5,6, 7, 9, 11, 12, 13, 14, 15, 17, and 18 to 48 were between 0.05 and 414nM.

Example 76 In Vivo Studies

The in vivo tests on acetylcholine-induced bronchospasm in guinea pigwere performed according to Konzett, H. and Rössler, F., Arch. Exp.Path. Pharmacol., vol. 195, pp. 71-74 (1940). Aqueous solutions of thetest compounds were instilled intratracheally in anaesthetisedmechanically ventilated guinea pigs. Bronchial response to intravenousacetylcholine challenge was determined before and after drugadministration and changes in pulmonary resistance at severaltime-points were expressed as percent of inhibition of bronchospasm. Thebronchodilator activity of the tested compounds persisted unchanged upto 24 hours after the administration.

Example 77 Plasma Stability of (Compound 1)

In order to demonstrate that the compounds are degraded, stability inrat and human plasma at 1 and 5 hours was tested for a representativecompound of the invention, which is compound 1. Briefly 10 μl of a stocksolution 250 μM of compound 1 in acetonitrile were added to 1 ml of ratand human plasma, and samples were incubated at 37° C. Plasma (50 μL)was taken after 0, 1 and 5 hours of incubation and added to 140 μl ofacetonitrile with addition of verapamil as internal standard (250ng/ml). Samples were analysed by HPLC-MS/MS analysis.

Plasma stability was calculated as percentage remaining after 1 and 5hours by dividing the peak area at 1 or 5 hours by the area of the peakat time 0. After 1 and 5 hours of incubation, less than 2% of compound 1was detected, indicating that compound 1 is very unstable in both plasmaspecies. The other compounds of the invention behave similarly.

Where a numerical limit or range is stated herein, the endpoints areincluded. Also, all values and subranges within a numerical limit orrange are specifically included as if explicitly written out.

Obviously, numerous modifications and variations of the presentinvention are possible in light of the above teachings. It is thereforeto be understood that, within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

All patents and other references mentioned above are incorporated infull herein by this reference, the same as if set forth at length.

The invention claimed is:
 1. A method for the treatment of abroncho-obstructive disease, comprising administering, to subject inneed thereof, an effective amount, of a salt represented by formula (I):

wherein: A is an optionally substituted aryl group, optionallysubstituted heteroaryl group, optionally substituted arylalkyl group,optionally substituted heteroarylalkyl group, or a group of formula (a)

wherein: R₃ and R₄ are the same or different and are independently H,(C₃-C₈)-cycloalkyl, aryl or heteroaryl, wherein said aryl or heteroarylmay be optionally substituted with a halogen atom or with one or moresubstituents independently selected from the group consisting of OH,O—(C₁-C₁₀)-alkyl, oxo, SH, S—(C₁-C₁₀)-alkyl, NO₂, CN, CONH₂, COOH,(C₁-C₁₀)-alkoxycarbonyl, (C₁-C₁₀)-alkylsulfanyl, (C₁-C₁₀)-alkylsulfinyl,(C₁-C₁₀)-alkylsulfonyl, (C₁-C₁₀)-alkyl, and (C₁-C₁₀)-alkoxyl or when R₃and R₄ are both independently aryl or heteroaryl they may be linkedthrough a Y group which is a (CH₂)_(n) group wherein n=0-2, wherein whenn=0, Y is a single bond, forming a tricyclic ring system wherein acarbon atom of (CH₂)_(n) may be substituted by heteroatom selected fromO, S, N, and with the proviso that R₃ and R₄ are never both H; R is agroup selected from the group consisting of: (C₁-C₁₀)-alkyl,(C₂-C₁₀)-alkenyl, and (C₂-C₁₀)-alkynyl each of which may be optionallysubstituted with a group selected from the group consisting of: NH₂,NR₁R₂, CONR₁R₂, NR₂COR₁, OH, SOR₁, SO₂R₁, SH, CN, NO₂, an alicycliccompound, Z—R₁, wherein Z is selected from CO, O, COO, OCO, SO₂, S, SO,COS, and SCO or it is a bond, and (C₃-C₈)-cycloalkyl; R₁ is a groupselected from the group consisting of: an alicyclic compound optionallysubstituted with one or more substituents independently selected fromthe group consisting of OH, oxo, SH, NO₂, CN, CONH₂,NR₂CO—(C₁-C_(x))-alkyl, COOH, (C₁-C₁₀)-alkoxycarbonyl,(C₁-C₁₀)-alkylsulfanyl, (C₁-C₁₀-alkylsulfinyl, (C₁-C₁₀)-alkylsulfonyl,(C₁-C₁₀)-alkyl, and (C₁-C₁₀)-alkoxyl NR₂CO—(C₁-C₁₀)-alkyl; an aryl groupoptionally substituted with NR₂CO—(C₁-C₁₀)-alkyl; and a heteroaryl groupoptionally substituted with a group selected from the group consistingof NR₂CO—(C₁-C₁₀)-alkyl, (C₁-C₁₀)-alkyl, O—(C₁-C₁₀)-alkyl, and halogen,R₂ is a group selected from the group consisting of H, phenoxycarbonyl,benzyloxycarbonyl, (C₁-C₁₀)-alkoxycarbonyl, (C₁-C₁₀)-alkylcarbonyl,(C₁-C₁₀)-alkylsulfonyl; and (C₁-C₁₀)-alkyl and X⁻ is a physiologicallyacceptable anion.
 2. A method according to claim 1, wherein saidbroncho-obstructive disease is asthma, chronic bronchitis, or chronicobstructive pulmonary disease.
 3. A method according to claim 1, whereinR is (C₁-C₆)-alkyl substituted by —Z—R₁.
 4. A method according to claim1, wherein R is (C₁-C₆)-alkyl substituted by —Z—R₁, wherein Z is O, CO,or a bond, and R₁ is aryl, optionally substituted with one or morehalogen atoms, or heteroaryl, optionally substituted with one or morehalogen atoms.
 5. A method according to claim 1, wherein R is CH₂—Z—R₁,wherein Z is CO and R₁ is thienyl.
 6. A method according to claim 5,wherein A is optionally substituted aryl, optionally substitutedheteroaryl, optionally substituted aryl alkyl, optionally substitutedheteroarylalkyl, or a group of formula (a) wherein R₃ and R₄ are bothindependently aryl, optionally substituted with one or more halogenatoms, or heteroaryl, optionally substituted with one or more halogenatoms.
 7. A method according to claim 5, wherein A is a group of formula(a) wherein R₃ and R₄ are both phenyl, optionally substituted with oneor more halogen atoms.
 8. A method according to claim 5, wherein A is agroup of formula (a) wherein R₃ and R₄ are both independently phenyl andthey are linked through a Y group which is a (CH₂)_(n) with n=0-2,wherein when n=0, Y is a single bond.
 9. A method according to claim 5,wherein A is a 9H-fluoren-9-yl group, optionally substituted with one ormore halogen atoms.
 10. A method according to claim 5, wherein A is abenzyl group, optionally substituted with one or more halogen atoms. 11.A method according to claim 5, wherein A is a biphenylmethyl group,optionally substituted with one or more halogen atoms.
 12. A methodaccording to claim 5, wherein A is a thiophenylmethyl group, optionallysubstituted with one or more halogen atoms.
 13. A method according toclaim 1, wherein R is a propyl group, substituted with a Z—R₁ group,wherein Z is O and R₁ is phenyl.
 14. A method according to claim 13,wherein A is aryl, heteroaryl, arylalkyl, heteroarylalkyl, or a group offormula (a) wherein R₃ and R₄ are both independently aryl, optionallysubstituted with one or more halogen atoms, or heteroaryl, optionallysubstituted with one or more halogen atoms.
 15. A method according toclaim 13, wherein A is a group of formula (a) wherein R₃ and R₄ are bothphenyl, optionally substituted with one or more halogen atoms.
 16. Amethod according to claim 13, wherein A is a group of formula (a)wherein R₃ and R₄ are both independently aryl or heteroaryl, and theyare linked through a Y group which is a (CH₂)_(n) with n=0-2, whereinwhen n=0, Y is a single bond.
 17. A method according to claim 13,wherein A is a 9H-fluoren-9-yl group, optionally substituted with one ormore halogen atoms.
 18. A method according to claim 13, wherein A is abenzyl group, optionally substituted with one or more halogen atoms. 19.A method according to claim 13, wherein A is a biphenylmethyl group,optionally substituted with one or more halogen atoms.
 20. A methodaccording to claim 13, wherein A is a thiophenylmethyl group, optionallysubstituted with one or more halogen atoms.